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The effects of L-arginine on protein stability and DNA binding ability of SaeR, a transcription factor in Staphylococcus aureus
Protein Expression and Purification ( IF 1.4 ) Pub Date : 2020-09-25 , DOI: 10.1016/j.pep.2020.105765
Ruochen Fan , Xian Shi , Binmei Guo , Jing Zhao , Jialu Liu , Chunshan Quan , Yuesheng Dong , Shengdi Fan

The SaeRS two-component system in Staphylococcus aureus controls the expression of a series of virulence factors, such as hemolysins, proteases, and coagulase. The response regulator, SaeR, belongs to the OmpR family with an N-terminal regulatory domain and a C-terminal DNA binding domain. To improve the production and stability of the recombinant protein SaeR, l-arginine (L-Arg) was added to the purification buffers. L-Arg enhanced the solubility and stability of the recombinant protein SaeR. The thermal denaturation temperature of SaeR in 10 mM L-Arg buffer was significantly increased compared to the buffer without L-Arg. Microscale Thermophoresis (MST) analysis results showed that the SaeR protein could bind to the P1 promoter under both phosphorylated and non-phosphorylated status in buffer containing 10 mM L-Arg. These results illustrate an effective method to purify SaeR and other proteins.



中文翻译:

L-精氨酸对金黄色葡萄球菌转录因子SaeR的蛋白质稳定性和DNA结合能力的影响

金黄色葡萄球菌中的SaeRS两组分系统控制一系列毒力因子的表达,例如溶血素,蛋白酶和凝固酶。响应调节剂SaeR属于OmpR家族,具有N端调节结构域和C端DNA结合结构域。为了提高重组蛋白SAER,生产和稳定性将-精氨酸(L-Arg)添加至纯化缓冲液。L-Arg增强了重组蛋白SaeR的溶解度和稳定性。与没有L-Arg的缓冲液相比,在10 mM L-Arg缓冲液中SaeR的热变性温度显着提高。微量热电泳(MST)分析结果表明,在含有10 mM L-Arg的缓冲液中,SaeR蛋白可以在磷酸化和非磷酸化状态下与P1启动子结合。这些结果说明了纯化SaeR和其他蛋白质的有效方法。

更新日期:2020-10-05
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